The overall objective is to elucidate the nature and mechanism(s) of action of those host and viral encoded/induced factors which regulate the transcription of both viral and cellular genes in human cells infected with adenovirus 2 and adenovirus 2-transformed cells. We have recently shown the accurate transcription of a variety of purified genes in cell systems reconstituted with purified RNA polymerases, crude extracts or partially purified factors, and purified DNA. These include adenovirus 2 early, intermediate, and late transcription units (encoding mRNAs and VA RNAs) as well as other viral and cellular transcription units (encoding mRNAs and tRNA and 5S RNAs). We have also identified several discrete host factors which are sufficient for the transcription of host and viral genes by polymerase III and discrete factors sufficient for the transcription of host and viral genes (encoding mRNAs) transcribed by polymerase II. In a further analysis of this problem we propose (1) to purify the various host factors to homogeneity, (2) to investigate their sites and mechanisms of action, including interactions with specific DNA sites, (3) to identify viral encoded/induced factors involved in the regulation of viral gene transcription, (4) to investigate the sites and mechanisms of action of these components, (5) as part of the preceding studies, to investigate the transcription of viral nucleoprotein templates (containing cellular histones or viral core proteins) and their possible role(s) in transcriptional regulation, and (6) as part of the preceding studies, to use site-specific mutagenesis and recombinant DNA technology to determine specific DNA sequences/nucleotides important for transcription.